石家庄地区肺炎支原体感染的流行病学调查

目的：了解石家庄地区肺炎支原体感染的血清流行病学情况。方法：选择2011年3月-2012年2月我院住院和门诊收治的急性呼吸道感染患者1902例为研究对象,采用间接免疫荧光法（IFA）检测其血清肺炎支原体IgM抗体,并分析其流行病学资料。结果：1902例血清标本中,284例（14．93％）肺炎支原体IgM抗体阳性,男性和女性的阳性率无显著差异。肺炎支原体抗体阳性的患者主要分布于0～15岁年龄段,阳性检出率最高的年龄组为0～6岁,占21．26％（132／621）。各个季节均有肺炎支原体感染阳性患者,感染率无显著差异性,秋（80例）、冬（96例）两季的阳性感染率高于春（56例）、夏（52例）两季。患者100％出现发热症状,95．77％出现咳嗽。结论：石家庄地区肺炎支原体感染的主要人群为未成年人,无季节性和性别差异,以发热和咳嗽为最主要的临床症状。     

基层中医院735例女性宫颈分泌物支原体检测及药敏分析

目的了解生殖道支原体感染与年龄、白带清洁度的关系及对抗生素的敏感性,为临床用药提供依据。方法对735例疑似生殖道感染标本采用珠海迪尔生物有限公司生产的试剂盒进行支原体培养和药敏试验,取阴道分泌物作清洁度检查。结果735例标本中检出支原体阳性402例,阳性率为54．7％,解脲脲原体（Uu）、Uu＋Mh混合感染及人型支原体（Mh）分别占75．1％、20．9％、4．0％。Uu、Uu＋Mh阳性患者好发于18～40岁。白带清洁度异常者的Uu、Uu＋Mh、Mh阳性率明显高于白带清洁度正常者。药敏结果显示,支原体对强力霉素、美满霉素敏感性较高,敏感率为85．7％～100．0％。结论白带清洁度异常者为支原体感染的高危人群,诊治时应常规做支原体检测,支原体感染的治疗应根据感染病原体的类型和药敏结果合理应用抗生素。     

550例支原体药敏结果分析

目的：了解本地区感染泌尿生殖系的解脲、人型支原体耐药情况。方法：对性病科和妇科门诊近3年来550例支原体感染者用支原体药敏试剂盒进行司巴沙星（SPA）、克拉霉素（CLA）、可乐必妥（CRA）、交沙霉素（JOS）、阿奇霉素（AZI）、罗红霉素（ROX）、强力霉素（DOX）、美满霉素（MIN）、氧氟沙星（ODL）、乙酰螺旋霉素（ASP）、四环素（TET）、红霉素（ERY）12种抗生素的药敏试验。结果：统计近3年耐药率,总计依次是4.2%、8.4%、10.4%、23.2%、23.8%、29.6%、29.6%、33.0%、43.1%、55.3%、61.9%、82.7%。结论：耐药率司巴沙星、克拉霉素、可乐必妥较稳定,基本低于10%,其余均较高且大部分呈逐年增长趋势,且解脲和人型支原体耐药性有较大差异。     

Syndrome with Joint Manifestations in Association with Mycoplasma pneumoniae Infection

Three patients experienced a rheumatic-fever-like illness with painful limitation of joint movements. The illnesses were due to infection with Mycoplasma pneumoniae. Joint symptoms have not been previously described in cases of M. pneumoniae infection.     

State of cellular immunity and phagocytes in guinea pigs infected with Mycoplasma pneumoniae

The work presents the results of the study of experimental Mycoplasma pneumoniae infection in guinea-pigs. A considerable increase in the engulfment of mycoplasmas and blood leukocytes was found to occur on days 14-28 after the infection. The correlation between the degree of the engulfment of mycoplasmas by macrophages and the activity of lymphocytes in the reaction of blast-cell transformation with phytohemagglutinin and Mycoplasma antigen was observed. The peculiarities of the course of infection in sensitized animals were revealed. Virulent Mycoplasma pneumoniae strains were found to have a toxic effect on the lymphocytes and macrophages of guinea-pigs. This effect was neutralized by specific antiserum. The importance of these facts for the pathogenesis of Mycoplasma infection is discussed.     

Simultaneous detection and identification of common cell culture contaminant and pathogenic mollicutes strains by reverse line blot hybridization

We have developed a reverse line blot (RLB) hybridization assay to detect and identify the commonest mollicutes causing cell line contamination (Mycoplasma arginini, Mycoplasma fermentans, Mycoplasma hyorhinis, Mycoplasma orale, and Acholeplasma laidlawii) and human infection (Mycoplasma pneumoniae, Mycoplasma hominis, Mycoplasma genitalium, Ureaplasma parvum, and Ureaplasma urealyticum). We developed a nested PCR assay with "universal" primers targeting the mollicute 16S-23S rRNA intergenic spacer region. Amplified biotin-labeled PCR products were hybridized to membrane-bound species-specific oligonucleotide probes. The assay correctly identified reference strains of 10 mollicute species. Cell cultures submitted for detection of mollicute contamination, clinical specimens, and clinical isolates were initially tested by PCR assay targeting a presumed mollicute-specific sequence of the 16S rRNA gene. Any that were positive were assessed by the RLB assay, with species-specific PCR assay as the reference method. Initially, 100 clinical and 88 of 92 cell culture specimens gave concordant results, including 18 in which two or more mollicute species were detected by both methods. PCR and sequencing of the 16S-23S rRNA intergenic spacer region and subsequent retesting by species-specific PCR assay of the four cell culture specimens for which results were initially discrepant confirmed the original RLB results. Sequencing of amplicons from 12 cell culture specimens that were positive in the 16S rRNA PCR assay but negative by both the RLB and species-specific PCR assays failed to identify any mollicute species. The RLB hybridization assay is sensitive and specific and able to rapidly detect and identify mollicute species from clinical and cell line specimens.     

支原体抑制宿主肿瘤细胞酪氨酸激酶受体c-kit降解

目的：研究支原体污染对宿主肿瘤细胞酪氨酸激酶受体c-kit降解的影响,探讨支原体污染对肿瘤细胞增殖的影响。方法：应用激光共聚焦荧光显微镜及透射电镜观察支原体污染HeLa细胞后内吞体的改变,间接反映外膜受体经内吞后的转运动力学改变：应用westernblot直接检测c-kit总量变化,泛素化水平及磷酸化水平;应用MTT法观察HeLa细胞污染支原体后细胞的增殖受到何种影响。结果：支原体污染后,内吞体聚集于核周,其膜表面积增大;支原体污染导致c—kit总量增加,同时泛素化磷酸化水平均增加,反映降解受抑制;支原体对肿瘤细胞增殖表现为双向影响,少量支原体污染促进HeLa细胞增殖,过量支原体污染则抑制增殖。结论：支原体污染在一定范围内可以通过抑制受体降解促进肿瘤细胞增殖,有可能是肿瘤进展中的一个加重因素,在将来需要引起临床工作者诊断和治疗上的重视。     

咽拭子快速培养在肺炎支原体感染中的临床应用价值

目的:探讨咽拭子快速培养在肺炎支原体感染中的临床应用价值。方法:收集2014年2月~2016年2月期间我院收治的呼吸道感染患儿220例,用肺炎支原体专用液体培养基进行肺炎支原体快速培养,用胶体金法检测肺炎支原体MP-Ig M。比较两种方法的阳性率。结果:咽拭子培养快速培养阳性率与血清MP-Ig M检测阳性率比较,差异无统计学意义(P0.05)。MP-Ig检测显示,≤1岁阳性率最低,其阳性率随年龄增加不断增高(P0.05)。肺炎支原体咽拭子培养显示,≤1岁阳性率最高,2~8岁最低(P0.05)。病程≤7 d患者肺炎支原体咽拭子培养阳性率(34.21%)显著高于肺炎支原体MP-Ig检测阳性率(14.04%)(P0.05)。病程7 d患者肺炎支原体咽拭子培养阳性率(11.32%)显著低于肺炎支原体MP-Ig检测阳性率(52.83%)(P0.05)。肺炎支原体咽拭子培养的灵敏度性以及特异性显著高于肺炎支原体MP-Ig检测,差异具有统计学意义(P0.05)。结论:咽拭子快速培养对肺炎支原体感染的早期诊断有一定临床应用价值,方法简单,无创伤,值得临床进一步研究和应用。     

A comparison of glycan expression and adhesion of mouse-adapted strains and clinical isolates of Helicobacter pylori

The role of Mycoplasma pneumoniae infection as a trigger for asthma exacerbations is well supported in previous studies. This study was designed to investigate the role of M. pneumoniae infection in acute exacerbation of asthma in children. A total of 150 patients (110 males, 40 females) were studied and immunoglobulin M (IgM) antibodies to M. pneumoniae were detected by enzyme-linked immunosorbent assay (ELISA), and PCR amplification was performed for the P1 gene to associate M. pneumoniae infection with asthma. As compared with 33 children with asthma, only two of the control subjects had positive IgM titers for M. pneumoniae , which was statistically significant ( P =0.002). A total of 15 children with asthma were positive by PCR for the P1 gene while none of the controls had a positive PCR. Of these positive cases, 24 cases were positive only by ELISA, six were positive only by PCR and nine patients were found to be positive by both ELISA and PCR. All the clinical characteristics of the patients at baseline were comparable between the moderate and the severe group of patients statistically, except for the peak expiratory flow rate. Mycoplasma pneumoniae infection was found to have a significant association with acute exacerbation in the moderate group of asthma patients by PCR ( P =0.01). These data suggest that M. pneumoniae infection may contribute to asthma exacerbation.     

杭州地区儿童肺炎支原体感染分析

目的调查杭州地区呼吸道感染儿童肺炎支原体（MP）感染情况和流行特点,为临床治疗和防止其爆发流行提供依据。方法随机采集在本院就诊并有呼吸道感染患儿的咽拭子,用FQ-PCR测定标本中MPDNA含量,若结果〉125拷贝为阳性,〈25拷贝为阴性。用统计学软件SPSS10．0分析MP感染和各调查因素之间的关系。结果在1502例呼吸道感染患儿标本中共检出391例MP阳性,阳性率占26．0％（391／1502）。感染机会、性别差异无显著性。7～15岁的儿童更容易感染,感染率占48．0％-62．1％。一年之中以夏秋季感染率最高,其中7月份高达37．3％。结论杭州地区MP引起儿童呼吸道感染的流行特点和南方其他地区的报道基本一致。     

女性生殖道炎症支原体多重耐药性分析

了解本地区在采用常规抗生素治疗支原体后多重耐药株的产生情况,为临床针对性用药提供科学依据。采用培养法对女性生殖道炎患者分泌物进行支原体培养、体外药敏试验及分析。结果可见,感染数量≥10~4 cfu/mL的支原体阳性率为52.9%(82/155);13种抗生素中,耐药种数构成比为6R>2R、4R、5R>3R>1R>8R>7R、10R>0R>9R、12R、13R;耐药率为林可霉素>壮观霉素>环丙沙星>诺氟沙星>司帕沙星>罗红霉素>氧氟沙星>克拉霉素>左氧氟沙星>阿齐霉素>强力霉素,美满霉素>交沙霉素。体外药敏试验对临床治疗支原体感染及减少多重耐药株的产生具有重要指导意义。     

Application of a Mycoplasma group-specific PCR for monitoring decontamination of Mycoplasma-infected Chlamydia sp. strains.

Mycoplasma contamination of biological materials remains a major problem. Most contaminations are caused by the use of Mycoplasma-contaminated cell lines. We adapted a Mycoplasma group-specific PCR to detect Mycoplasma contamination in cell lines and demonstrate its use in monitoring decontamination procedures with Mycoplasma-contaminated suspensions of Chlamydia spp. Three different methods were investigated: the use of Mycoplasma-specific antiserum in cell culture, physical separation by the combined use of enzymatic treatment and differential centrifugation, and the use of detergents. With these methods only incubation with Triton X-100 resulted in decontamination of Mycoplasma-contaminated suspensions of several laboratory strains of Chlamydia pneumoniae, C. pecorum, and C. trachomatis. Only one C. pneumoniae strain, UZG-1, was sensitive to Triton X-100 treatment. Since 39 of 40 throat swabs from patients with symptoms of an upper respiratory tract infection had positive reactions in the Mycoplasma group-specific PCR, this procedure could also have clinical significance in attempts to propagate C. pneumoniae strains from clinical specimens.     

Inactivated vaccine induces protection against Mycoplasma agalactiae infection in sheep

The efficacy of an inactivated oil-emulsion vaccine against Mycoplasma agalactiae was evaluated by an experimental infection of sheep. The vaccinated sheep developed high levels of antibodies and, following challenge, they did not develop any clinical signs of disease and the mycoplasmas were not detected, either by isolation trials or PCR assays carried out both on nasal swabs and milk specimens. The unvaccinated-challenged sheep showed typical signs of M. agalactiae infection and bacterial shedding. The results obtained indicate a good efficacy of the vaccine in eliciting protection against M. agalactiae infection.     

女性泌尿生殖道支原体感染及耐药性分析

目的通过分析深圳市人民医院2009年9月至2010年4月女性泌尿生殖道支原体感染状况及耐药性变迁,指导临床合理用药。方法采用法国生物梅里埃公司MyeoplasmaIST2支原体鉴定及药敏试剂盒,进行支原体检测及药敏分析。结果2135例患者总检出率为54．6％（1166／2135）,其中解脲脲支原体（Uu）阳性1017例（47．6％）;人型支原体（Mh）阳性32例（1．5％）;Uu＋Mh阳性117例（5．5％）。1166例患者支原体对9种抗生素够感性依次为原始霉素（99．8％）、交沙霉素（99．6％）、强力霉素（96．3％）、四环素（94．4％）、克拉霉素（85．6％）、阿奇霉素（77．6％）、红霉素（75．6％）、氧氟沙星（28．4％）和环丙沙星（22．8％）。结论支原体（尤其是Uu）感染已成为女性泌尿生殖系统炎症的主要病原体之一,治疗支原体感染应选择原始霉素、交沙霉素、强力霉素等敏感率高的药物及药敏试验显示敏感的药物,喹诺酮类药物耐药率较高,应引起临床重视。     

Simultaneous Detection and Identification of Common Cell Culture Contaminant and Pathogenic Mollicutes Strains by Reverse Line Blot Hybridization

We have developed a reverse line blot (RLB) hybridization assay to detect and identify the commonest mollicutes causing cell line contamination (Mycoplasma arginini, Mycoplasma fermentans, Mycoplasma hyorhinis, Mycoplasma orale, and Acholeplasma laidlawii) and human infection (Mycoplasma pneumoniae, Mycoplasma hominis, Mycoplasma genitalium, Ureaplasma parvum, and Ureaplasma urealyticum). We developed a nested PCR assay with “universal” primers targeting the mollicute 16S-23S rRNA intergenic spacer region. Amplified biotin-labeled PCR products were hybridized to membrane-bound species-specific oligonucleotide probes. The assay correctly identified reference strains of 10 mollicute species. Cell cultures submitted for detection of mollicute contamination, clinical specimens, and clinical isolates were initially tested by PCR assay targeting a presumed mollicute-specific sequence of the 16S rRNA gene. Any that were positive were assessed by the RLB assay, with species-specific PCR assay as the reference method. Initially, 100 clinical and 88 of 92 cell culture specimens gave concordant results, including 18 in which two or more mollicute species were detected by both methods. PCR and sequencing of the 16S-23S rRNA intergenic spacer region and subsequent retesting by species-specific PCR assay of the four cell culture specimens for which results were initially discrepant confirmed the original RLB results. Sequencing of amplicons from 12 cell culture specimens that were positive in the 16S rRNA PCR assay but negative by both the RLB and species-specific PCR assays failed to identify any mollicute species. The RLB hybridization assay is sensitive and specific and able to rapidly detect and identify mollicute species from clinical and cell line specimens.     

大学生尿沉渣中解脲脲原体和人型支原体感染情况分析

目的了解解脲脲原体(Ureaplasma urealytium,Uu)和人型支原体(Mycoplasma homanis,Mh)在大学生泌尿道中的感染情况.方法采用培养方法对37例正常的在校大学生的尿沉渣进行Uu和Mh的培养.结果采用培养方法检测21.6%(8/37)的大学生Uu感染为阳性,其中分离阳性者全部是女生(8/20,40%),13.50%(5/37)大学生Mh培养为阳性,其中阳性者亦全部为女生,占女生人数的25%(5/20).结论Uu和Mh可能是女性泌尿道的正常寄生的微生物之一.     

High prevalence of Mycoplasma infections among European chronic fatigue syndrome patients. Examination of four Mycoplasma species in blood of chronic fatigue syndrome patients

Prevalence of Mycoplasma species infections in chronic fatigue syndrome (CFS) has been extensively reported in the scientific literature. However, all previous reports highlighted the presence of Mycoplasmas in American patients. In this prospective study, the presence of Mycoplasma fermentans, M. penetrans, M. pneumoniae and M. hominis in the blood of 261 European CFS patients and 36 healthy volunteers was examined using forensic polymerase chain reaction. One hundred and seventy-nine (68.6%) patients were infected by at least one species of Mycoplasma, compared to two out of 36 (5.6%) in the control sample (P<0.001). Among Mycoplasma-infected patients, M. hominis was the most frequently observed infection (n=96; 36.8% of the overall sample), followed by M. pneumoniae and M. fermentans infections (equal frequencies; n=67; 25.7%). M. penetrans infections were not found. Multiple mycoplasmal infections were detected in 45 patients (17.2%). Compared to American CFS patients (M. pneumoniae>M. hominis>M. penetrans), a slightly different pattern of mycoplasmal infections was found in European CFS patients (M. hominis>M. pneumoniae, M. fermentansz.Gt;M. penetrans).     

泌尿生殖道支原体感染检测及药敏试验

目的了解本地区泌尿生殖道解脲脲原体（Uu）和人型支原体（Mh）感染状况及药物敏感性,指导临床医生合理应用抗生素。方法应用法国生物梅里埃公司提供的IST试剂盒进行支原体鉴定及9种药物敏感检测,并对结果进行统计分析。结果1210例门诊患者检出支原体阳性683例,总感染率为56.4%,其中Uu单独感染占628例（占51.9%）,Mh单独感染14例（占1.2%）,Uu和Mh混合感染41例（占3.4%）。交沙霉素和原始霉素敏感率最高,对Uu分别为98.5%和97.0%,对Uu和Mh混合感染率都为100%;氧氟沙星敏感率最低,分别为1.5%和0.0%。结论泌尿生殖道系统感染主要由解脲支原体引起,交沙霉素和原始霉素敏感率最高,氧氟沙星敏感率最低。临床应选用培养敏感的抗菌药物,提高治疗效果。     

肺炎支原体感染小鼠体内IL-6、IFN-γ、IL-5的水平及克拉霉素的影响

目的：研究肺炎支原体感染小鼠血清及支气管肺泡灌洗液中细胞因子的水平,了解克拉霉素治疗对细胞因子的影响。方法：将昆明小鼠分为感染组、正常组及药物治疗组,建立小鼠肺炎支原体感染模型成功后,药物组用克拉霉素（6g·kg^-1·d^-1）进行治疗,连续5天。在肺炎支原体感染模型建立后的第8天,检测三组小鼠的血清及支气管肺泡灌洗液中IL-6、IFN-γ、IL-5水平。结果-相比较于正常组,感染组小鼠支气管灌洗液和血清中的IL-6、IFN-γ、IL-5均显著升高（P〈0．05）。而克拉霉素的使用能使IL-6、IFN-γ降低（P〈0．05）,但对IL-5水平无影响。结论：肺炎支原体感染使小鼠IL-6、IFN-γ、IL-5水平增高,克拉霉素治疗有一定疗效。     

儿科呼吸道疾病中肺炎支原体感染的结果分析

目的探讨儿科肺炎支原体（MP）感染特点,辅助临床医师早期诊断,合理用药。方法测定我院一年来2013例儿科呼吸道疾病患儿的肺炎支原体抗体（IgM）。结果2013例呼吸道感染患儿,检出肺炎支原体抗体（IgM）阳性者769例,占38.2%,769例阳性分别表现为肺炎369例（48%）,支气管炎238例（31%）,咽炎92例（12%）,哮喘70例（9%）。其中,肺炎组与各组相比较,具有统计学意义。MP IgM感染的检出率明显高于其他各组（P〈0.01）。结论MP感染是患儿不可忽视的病原体,检测患儿血清MP抗体能够及早诊断,指导治疗。